Publications by authors named "Tianlai Li"

49 Publications

Genome-Wide Characterization of B-Box Gene Family and Its Roles in Responses to Light Quality and Cold Stress in Tomato.

Front Plant Sci 2021 5;12:698525. Epub 2021 Jul 5.

College of Horticulture, Shenyang Agricultural University, Shenyang, China.

Perceiving incoming environmental information is critical for optimizing plant growth and development. Multiple B-box proteins (BBXs) play essential roles in light-dependent developmental processes in plants. However, whether BBXs function as a signal integrator between light and temperature in tomato plants remains elusive. In this study, 31 genes were identified from the newly released tomato () genome sequences and were clustered into five subgroups. Gene structure and protein motif analyses showed relatively high conservation of closely clustered genes within each subgroup; however, genome mapping analysis indicated the uneven distribution of the genes on tomato chromosomes. Promoter -regulatory elements prediction and gene expression indicated that genes were highly responsive to light, hormones, and stress conditions. Reverse genetic approaches revealed that disruption of , and largely suppressed the cold tolerance of tomato plants. Furthermore, the impairment of , and suppressed the photosynthetic response immediately after cold stress. Due to the impairment of non-photochemical quenching (NPQ), the excess photon energy and electron flow excited by low temperature were not consumed in , and silenced plants, leading to the over reduction of electron carriers and damage of the photosystem. Our study emphasized the positive roles of light signaling transcription factors SlBBXs in cold tolerance in tomato plants, which may improve the current understanding of how plants integrate light and temperature signals to adapt to adverse environments.
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http://dx.doi.org/10.3389/fpls.2021.698525DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8287887PMC
July 2021

Effect of the ridge position ratio on the thermal environment of the Chinese solar greenhouse.

R Soc Open Sci 2021 May 12;8(5):201707. Epub 2021 May 12.

College of Engineering, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, Shenyang 110866, People's Republic of China.

This paper clarified the mechanism of the south and north roofs' effect on the thermal environment of the Chinese solar greenhouse (CSG), using a new parameter: ridge position ratio (RPR), which can describe the dynamic dependency relationship between the south and north roofs. A mathematical model was established using a method of combining computational fluid dynamics (CFD) simulation with experiments, then the model was used to further analyse the effect of RPR on the thermal environment of the CSG. The experimental greenhouse was simulated as an empty building to obtain results independently from these factors including crop and ventilation conditions. The results showed that the occurrence time of the maximum air temperature will be delayed when RPR increases to 0.3 during the daytime. As RPR increases, the heat storage layer of the soil gradually becomes thinner, but the north wall remains unchanged. RPR has a relatively small effect on the minimum temperature of each greenhouse part during the night. Mathematical models of the relationships between RPR, the solar energy that entered the greenhouse and the released heat energy of the enclosure structures were established, respectively. This paper can provide theoretical guidance for the structural design of the CSG.
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http://dx.doi.org/10.1098/rsos.201707DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113901PMC
May 2021

Inflorescence abscission protein SlIDL6 promotes low light intensity-induced tomato flower abscission.

Plant Physiol 2021 Mar 12. Epub 2021 Mar 12.

College of Horticulture, Shenyang Agricultural University, Shenyang, Liaoning 110866, People's Republic of China.

In many fruiting plant species, flower abscission is induced by low light stress. Here, we elucidated how signaling mediated by the peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) controls low light-induced flower drop in tomato (Solanum lycopersicum). We analyzed the expression patterns of an IDA-Like gene (SlIDL6) during low light-induced flower abscission, and used tandem mass spectrometry to identify and characterize the mature SlIDL6 peptide. Tomato knockout lines were created to investigate the in vivo function of SlIDL6. In addition, yeast one-hybrid assays were used to investigate the binding of the SlWRKY17 transcription factor to the SlIDL6 promoter, and silencing of SlWRKY17 expression delayed low light-induced flower abscission. SlIDL6 was specifically expressed in the abscission zone (AZ) and at high levels during low light-induced abscission and ethylene treatment. SlIDL6 knockout lines showed delayed low light-induced flower drop, and the application of SlIDL6 peptide accelerated abscission. Overexpression of SlIDL6 rescued the ida mutant phenotype in Arabidopsis (Arabidopsis thaliana), suggesting functional conservation between species. SlIDL6-mediated abscission was via an ethylene-independent pathway. We report a SlWRKY17- SlIDL6 regulatory module that functions in low light promoted abscission by increasing the expression of enzymes involved in cell wall remodeling and disassembly.
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http://dx.doi.org/10.1093/plphys/kiab120DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8195530PMC
March 2021

Inflorescence abscission protein SlIDL6 promotes low light intensityinduced tomato flower abscission.

Plant Physiol 2021 Mar 12. Epub 2021 Mar 12.

College of Horticulture, Shenyang Agricultural University, Shenyang, Liaoning 110866, People's Republic of China.

In many fruiting plant species, flower abscission is induced by low light stress. Here, we elucidated how signaling mediated by the peptide INFLORESCENCE DEFICIENT IN ABSCISSION (IDA) controls low light-induced flower drop in tomato (Solanum lycopersicum). We analyzed the expression patterns of an IDA-Like gene (SlIDL6) during low light-induced flower abscission, and used tandem mass spectrometry to identify and characterize the mature SlIDL6 peptide. Tomato knockout lines were created to investigate the in vivo function of SlIDL6. In addition, yeast one-hybrid assays were used to investigate the binding of the SlWRKY17 transcription factor to the SlIDL6 promoter, and silencing of SlWRKY17 expression delayed low light-induced flower abscission. SlIDL6 was specifically expressed in the abscission zone (AZ) and at high levels during low light-induced abscission and ethylene treatment. SlIDL6 knockout lines showed delayed low light-induced flower drop, and the application of SlIDL6 peptide accelerated abscission. Overexpression of SlIDL6 rescued the ida mutant phenotype in Arabidopsis (Arabidopsis thaliana), suggesting functional conservation between species. SlIDL6-mediated abscission was via an ethylene-independent pathway. We report a SlWRKY17- SlIDL6 regulatory module that functions in low light promoted abscission by increasing the expression of enzymes involved in cell wall remodeling and disassembly.
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http://dx.doi.org/10.1093/plphys/kiab121DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8195514PMC
March 2021

SlERF52 regulates SlTIP1;1 expression to accelerate tomato pedicel abscission.

Plant Physiol 2021 Apr;185(4):1829-1846

College of Horticulture, Shenyang Agricultural University, Shenyang, Liaoning 110866, People's Republic of China.

Abscission of plant organs is induced by developmental signals and diverse environmental stimuli and involves multiple regulatory networks, including biotic or abiotic stress-impaired auxin flux in the abscission zone (AZ). Depletion of auxin activates AZ ethylene (ETH) production and triggers acceleration of abscission, a process that requires hydrogen peroxide (H2O2). However, the interaction between these networks and the underlying mechanisms that control abscission are poorly understood. Here, we found that expression of tonoplast intrinsic proteins, which belong to the aquaporin (AQP) family in the AZ was important for tomato (Solanum lycopersicum) pedicel abscission. Liquid chromatography-tandem mass spectrometry and in situ hybridization revealed that SlTIP1;1 was most abundant and specifically present in the tomato pedicel AZ. SlTIP1;1 localized in the plasma membrane and tonoplast. Knockout of SlTIP1;1 resulted in delayed abscission, whereas overexpression of SlTIP1;1 accelerated abscission. Further analysis indicated that SlTIP1;1 mediated abscission via gating of cytoplasmic H2O2 concentrations and osmotic water permeability (Pf). Elevated cytoplasmic levels of H2O2 caused a suppressed auxin signal in the early abscission stage and enhanced ETH production during abscission. Furthermore, we found that increasing Pf was required to enhance the turgor pressure to supply the break force for AZ cell separation. Moreover, we observed that SlERF52 bound directly to the SlTIP1;1 promoter to regulate its expression, demonstrating a positive loop in which cytoplasmic H2O2 activates ETH production, which activates SlERF52. This, in turn, induces SlTIP1;1, which leads to elevated cytoplasmic H2O2 and water influx.
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http://dx.doi.org/10.1093/plphys/kiab026DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8133580PMC
April 2021

Exogenous DA-6 Improves the Low Night Temperature Tolerance of Tomato Through Regulating Cytokinin.

Front Plant Sci 2020 4;11:599111. Epub 2021 Feb 4.

College of Horticulture, Shenyang Agricultural University, Shenyang, China.

Low night temperature (LNT) causes environmental stress and has a severe and negative impact on plant growth and productivity. Synthetic elicitors can regulate plant growth and induce defense mechanisms from this type of stress. Here, we evaluated the effect of the exogenous growth regulator diethyl aminoethyl hexanoate (DA-6) in tomato leaf response to LNT stress. Our results showed that exogenous DA-6 activates the expression of chlorophyll synthesis and photosystem-related genes, and results in higher photosynthetic activity and chlorophyll production. Furthermore, DA-6 can regulate the synthesis of endogenous cytokinin (CTK) and the expression of decomposition genes to stabilize chloroplast structure, reduce oxidative damage, and maintain the photochemical activity of tomato leaves under LNT stress. DA-6 maintains a high level of ABA content and induces the expression of genes, indicating that DA-6 may participate in the cold response signaling pathway and induce the expression of downstream low temperature response genes and accumulation of compatible osmolytes. This study unravels a mode of action by which plant growth regulators can improve low temperature tolerance and provides important considerations for their application to alleviate the harmful effects of cold stress.
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http://dx.doi.org/10.3389/fpls.2020.599111DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7889814PMC
February 2021

Correction to: miR168 targets Argonaute1A mediated miRNAs regulation pathways in response to potassium deficiency stress in tomato.

BMC Plant Biol 2021 Feb 8;21(1):84. Epub 2021 Feb 8.

Horticulture Department, College of Horticulture, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, Shenyang, 110866, P. R. China.

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http://dx.doi.org/10.1186/s12870-021-02855-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7871378PMC
February 2021

Exogenous Calcium Alleviates Nocturnal Chilling-Induced Feedback Inhibition of Photosynthesis by Improving Sink Demand in Peanut ().

Front Plant Sci 2020 21;11:607029. Epub 2020 Dec 21.

The UWA Institute of Agriculture, The University of Western Australia, Perth, WA, Australia.

(peanut) is a globally important oilseed crop with high nutritional value. However, upon exposure to overnight chilling stress, it shows poor growth and seedling necrosis in many cultivation areas worldwide. Calcium (Ca) enhances chilling resistance in various plant species. We undertook a pot experiment to investigate the effects of exogenous Ca and a calmodulin (CaM) inhibitor on growth and photosynthetic characteristics of peanut exposed to low night temperature (LNT) stress following warm sunny days. The LNT stress reduced growth, leaf extension, biomass accumulation, gas exchange rates, and photosynthetic electron transport rates. Following LNT stress, we observed larger starch grains and a concomitant increase in nonstructural carbohydrates and hydrogen peroxide (HO) concentrations. The LNT stress further induced photoinhibition and caused structural damage to the chloroplast grana. Exogenous Ca enhanced plant growth following LNT stress, possibly by allowing continued export of carbohydrates from leaves. Foliar Ca likely alleviated the nocturnal chilling-dependent feedback limitation on photosynthesis in the daytime by increasing sink demand. The foliar Ca pretreatment protected the photosystems from photoinhibition by facilitating cyclic electron flow (CEF) and decreasing the proton gradient (pH) across thylakoid membranes during LNT stress. Foliar application of a CaM inhibitor increased the negative impact of LNT stress on photosynthetic processes, confirming that Ca-CaM played an important role in alleviating photosynthetic inhibition due to the overnight chilling-dependent feedback.
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http://dx.doi.org/10.3389/fpls.2020.607029DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7779555PMC
December 2020

Effects of orientation and structure on solar radiation interception in Chinese solar greenhouse.

PLoS One 2020 6;15(11):e0242002. Epub 2020 Nov 6.

College of Horticulture, Shenyang Agricultural University, Shenyang, China.

In order to further improve the utilization of solar energy in Chinese Solar Greenhouse (CSG), this paper systematically studied the effects of orientation and structure on solar radiation interception in CSG. A solar radiation model has been developed based on the previous research, which taking solar motion law, meteorological data, and optical properties of materials into consideration. The established model was used to optimize the orientation and structure of CSG. The analysis of structure considered two major structural parameters, which are the ridge height and the horizontal projection of the rear roof. Moreover, the widely used Liao-Shen type Chinese solar greenhouse (CSG-LS) has been taken as the prototype in the present research, and the measured data of the typical clear day was used for the model validation. The results showed that the ridge height has a remarkable influence on the solar energy captured by CSG-LS. Compared with the optimization of a single factor, the comprehensive optimization of orientation and structure can increase the solar radiation interception of the rear wall by 3.95%. Considering the limiting factor of heat storage-release capacity and the shading effect on the greenhouse structure, the optimal lighting construction of the CSG-LS (with a span of 9.0 m) was specified as 7~9° from south to west of azimuth angle, 4.5~4.7 m ridge height, and 1.4~1.6 m horizontal projection of the rear roof at 42°N latitude. The proposed solar radiation model can provide scientific guidance for the CSG-LS construction in different areas.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0242002PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7647090PMC
December 2020

miR168 targets Argonaute1A mediated miRNAs regulation pathways in response to potassium deficiency stress in tomato.

BMC Plant Biol 2020 Oct 19;20(1):477. Epub 2020 Oct 19.

Horticulture Department, College of Horticulture, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, Shenyang, 110866, P.R. China.

Background: Potassium (K) is an essential ion for most plants, as it is involved in the regulation of growth and development. K homeostasis in plant cells has evolved to facilitate plant adaptation to K-deficiency stress. Argonaute1 (AGO1) is regulated by miR168 to modulate the small RNA regulatory pathway by RNA silencing complex (RISC) in tomatoes. However, the role of miR168-mediated regulation of AGO1 in the context of K deficiency stress in tomatoes has not been elucidated yet.

Results: SlmiR168 and its target gene SlAGO1A were differentially expressed among low-K-tolerant JZ34 and low-K-sensitive JZ18 tomato plants. Transgenic tomato plants constitutively expressing pri-SlmiR168a showed stronger root system growth, better leaves development, and higher K contents in roots under K-deficiency stress than those of the transgenic tomato lines expressing rSlAGO1A (SlmiR168-resistant) and the wild type (WT). Deep sequencing analysis showed that 62 known microRNAs (miRNAs) were up-regulated in 35S:rSlAGO1 compared with WT tomatoes. The same miRNAs were down-regulated in 35S:SlmiR168a compared with WT plants. The integrated analysis found 12 miRNA/mRNA pairs from the 62 miRNAs, including the root growth and cytokinin (CTK)/abscisic acid (ABA) pathways.

Conclusions: The regulation mediated by SlmiR168 of SlAGO1A contributes to the plant development under low-K stress. Moreover, this regulation mechanism may influence downstream miRNA pathways in response to low-K stress through the CTK/ABA and root growth modulation pathways.
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http://dx.doi.org/10.1186/s12870-020-02660-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7574427PMC
October 2020

Evaluation of airflow pattern and thermal behavior of the arched greenhouses with designed roof ventilation scenarios using CFD simulation.

PLoS One 2020 29;15(9):e0239851. Epub 2020 Sep 29.

National & Local Joint Engineering Research Center of Northern Horticultural Facilities Design & Application Technology (Liaoning), Shenyang, China.

Natural ventilation is an effective energy-saving strategy conducive to promoting sustainable agricultural production. A comprehensive numerical simulation is performed to predict the airflow pattern and thermal behavior in different arched greenhouses. The defined arc chord angle and position angle are employed to examine the natural ventilation process and corresponding roof vent scenarios. The numerical simulation is compared with the experimental data and good agreements are observed. Various configurations of ventilated structures, wind conditions and ventilation layouts are simulated on a high-resolution polyhedral grid based on a grid sensitivity analysis, which is beneficial to the optimization of greenhouse cooling combined with the water circulation heat collection system. The cooling effect in summer is analyzed by estimating the ventilation flow rate and microclimate inhomogeneity. The results demonstrate that the position angle of 85° of the arched greenhouses is an optimum ventilation direction and its impact on the microclimate is marginally affected by the change of the ventilation structure. The designed ventilation scheme has perfect air exchange capacity and cooling effect because the average air temperature can be reduced by 1.5°C more than the existing greenhouse in 10 minutes of ventilation. Likewise, the results show that the temperature and velocity inhomogeneities are approximately decreased by 33.3% and 11.89%, respectively. The practical value of the research is expected to provide basic quantitative conclusions for evaluating the natural ventilation performance.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0239851PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7523980PMC
November 2020

Tomato SlIDA has a critical role in tomato fertilization by modifying reactive oxygen species homeostasis.

Plant J 2020 09 11;103(6):2100-2118. Epub 2020 Jul 11.

College of Horticulture, Shenyang Agricultural University, Shenyang, Liaoning, 110866, People's Republic of China.

Anther development and pollen tube elongation are key steps for pollination and fertilization. The timing and spatial distribution of reactive oxygen species (ROS) and programmed cell death are central to these processes, but the regulatory mechanism of ROS production is not well understood. Inflorescence deficient in abscission (IDA) is implicated in many plant development and responses to environmental stimuli. However, their role in reproductive development is still unknown. We generated tomato knockout lines (CR-slida) of an IDA homolog (SlIDA), which is expressed in the tapetum, septum and pollen tube, and observed a severe defect in male gametes. Further analysis indicated that there was a programmed cell death defect in the tapetum and septum and a failure of anther dehiscence in the CR-slida lines, likely related to insufficient ROS signal. Liquid chromatography-tandem mass spectrometry identified mature SlIDA as a 14-mer EPIP peptide, which was shown to be secreted, and a complementation experiment showed that application of a synthetic 14-mer EPIP peptide rescued the CR-slida defect and enhanced the ROS signal. Moreover, the application of the ROS scavengers diphenyleneiodonium or Mn-TMPP suppressed peptide function. Collectively, our results revealed that SlIDA plays an essential role in pollen development and pollen tube elongation by modulating ROS homeostasis.
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http://dx.doi.org/10.1111/tpj.14886DOI Listing
September 2020

Tomato YABBY2b controls plant height through regulating indole-3-acetic acid-amido synthetase (GH3.8) expression.

Plant Sci 2020 Aug 27;297:110530. Epub 2020 May 27.

Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District 110866, PR China; National & Local Joint Engineering Research Center of Northern Horticultural Facilities Design & Application Technology (Liaoning), No. 120 Dongling Road, Shenhe District 110866, PR China; Key Laboratory of Protected Horticulture (Shenyang Agricultural University), Ministry of Education, No. 120 Dongling Road, Shenhe District 110866, PR China. Electronic address:

Dwarfing is a desirable agronomic trait in cultivation management. Dwarf plants are lodging-resistant, compact, and perform well under high-density planting. The use of dwarf genetic resources is one approach to improve crop yield. YABBY2b in tomato (Solanum lycopersicum) encodes a transcription factor that regulates plant height. In this study, we created YABBY2b knockout mutant lines, and the resulting yabby2b plants exhibited reduced height and smaller flowers and fruits. The RNA-seq analysis showed that 17 genes responding to gibberellin and auxin were differentially expressed. We hypothesized that indole-3-acetic acid-amido synthetase GH3.8 (GH3.8) played a crucial role in the resulting yabby2b dwarf phenotype. Further analysis showed that YABBY2b suppresses GH3.8 gene expression by directly binding to its promoter, and that this contributes to auxin-mediated repression of GH3.8. Moreover, the silencing of GH3.8 led to increased plant height. Combined, our data suggest that YABBY2b may positively regulate plant height in tomato by inhibiting the expression of growth suppressor GH3.8.
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http://dx.doi.org/10.1016/j.plantsci.2020.110530DOI Listing
August 2020

PGR5/PGRL1 and NDH Mediate Far-Red Light-Induced Photoprotection in Response to Chilling Stress in Tomato.

Front Plant Sci 2020 27;11:669. Epub 2020 May 27.

College of Horticulture, Shenyang Agricultural University, Shenyang, China.

Plants experience low ambient temperature and low red to far-red ratios (L-R/FR) of light due to vegetative shading and longer twilight durations in cool seasons. Low temperature induce photoinhibition through inactivation of the photosynthetic apparatus, however, the role of light quality on photoprotection during cold stress remains poorly understood. Here, we report that L-R/FR significantly prevents the overreduction of the entire intersystem electron transfer chain and the limitation of photosystem I (PSI) acceptor side, eventually alleviating the cold-induced photoinhibition. During cold stress, L-R/FR activated cyclic electron flow (CEF), enhanced protonation of PSII subunit S (PsbS) and de-epoxidation state of the xanthophyll cycle, and promoted energy-dependent quenching (qE) component of non-photochemical quenching (NPQ), enzyme activity of Foyer-Halliwell-Asada cycle and D1 proteins accumulation. However, L-R/FR -induced photoprotection pathways were compromised in tomato () and () co-silenced plants and () -silenced plants during cold stress. Our results demonstrate that both PGR5/PGRL1- and NDH-dependent CEF mediate L-R/FR -induced cold tolerance by enhancing the thermal dissipation and the repair of photodamaged PSII, thereby mitigating the overreduction of electron carriers and the accumulation of reactive oxygen species. The study indicates that there is an anterograde link between photoreception and photoprotection in tomato plants during cold stress.
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http://dx.doi.org/10.3389/fpls.2020.00669DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7270563PMC
May 2020

Effect of internal surface structure of the north wall on Chinese solar greenhouse thermal microclimate based on computational fluid dynamics.

PLoS One 2020 15;15(4):e0231316. Epub 2020 Apr 15.

College of Horticulture, Shenyang Agricultural University, Shenyang, China.

Chinese solar greenhouses are unique facility agriculture buildings and widely used in northeastern China, providing a favorable requirement for crop growth. The north wall configurations play an essential role in heat storage and thermal insulation and directly affect the management of the internal environment. This research is devoted to further improve the thermal performance of the greenhouse and explore the potential of the north wall. A mathematical model was designed to investigate the concave-convex wall configurations based on computational fluid dynamics. Four passive heat-storage north walls were analyzed by using the same constituent materials, including a plane wall, a vertical wall, a horizontal wall and an alveolate wall. The numerical model was validated by experimental measurements. The temperature distributions of the north walls were examined and a comparative analysis of the heat storage-release capabilities was carried out. The results showed that the heat-storage capacity of the north wall is affected by the surface structure. Moreover, the critical factor influencing the air temperature is the sum of the heat load released by the wall and the energy increment of greenhouse air. The results suggested that the alveolate wall has preferable thermal accumulation capacity. The concave-convex wall configurations have a wider range of heat transfer performance along the thickness direction, while the plane wall has a superior thermal environment. This study provides a basic theoretical reference to rationally design the internal surface structures of the north wall.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0231316PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7159206PMC
July 2020

Photosynthetic Response Mechanism of Soil Salinity-Induced Cross-Tolerance to Subsequent Drought Stress in Tomato Plants.

Plants (Basel) 2020 Mar 16;9(3). Epub 2020 Mar 16.

Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District 110866, China.

Soil salinization and water shortage cause ion imbalance and hyperosmoticity in plant cells, adversely impairing photosynthesis efficiency. How soil salinity-induced photosynthetic acclimation influences the cross-tolerance to drought conditions represents a promising research topic. This study was to reveal the photosynthetic mechanism of soil salinity-induced resistance to the subsequent drought stress in tomato leaves through comprehensive photosynthesis-related spectroscopy analysis. We conducted soil salinity pretreatment and subsequent drought stress experiments, including irrigation with 100 mL water, 100 mL 100 mM NaCl solution (NaCl100), 50 mL water, and 50 mL 100 mM NaCl solution (NaCl50) for five days, followed by five-day drought stress. The results showed that soil salinity treatment by NaCl decreased the rate of photosynthetic gas exchange but enhanced CO assimilation, along with photosystem II [PS(II)] and photosystem I [PS(I)] activity and photochemical efficiency in tomato plants compared with water pretreatment after subsequent drought stress. NaCl100 and NaCl50 had the capacity to maintain non-photochemical quenching (NPQ) of chlorophyll fluorescence and the cyclic electron (CEF) flow around PSI in tomato leaves after being subjected to subsequent drought stress, thus avoiding the decrease of photosynthetic efficiency under drought conditions. NaCl100 and NaCl50 pretreatment induced a higher proton motive force (pmf) and also alleviated the damage to the thylakoid membrane and adenosine triphosphate (ATP) synthase of tomato leaves caused by subsequent drought stress. Overall, soil salinity treatment could enhance drought resistance in tomato plants by inducing NPQ, maintaining CEF and pmf that tradeoff between photoprotection and photochemistry reactions. This study also provides a photosynthetic perspective for salt and drought cross-tolerance.
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http://dx.doi.org/10.3390/plants9030363DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7154942PMC
March 2020

Supplementary Calcium Restores Peanut () Growth and Photosynthetic Capacity Under Low Nocturnal Temperature.

Front Plant Sci 2019 21;10:1637. Epub 2020 Jan 21.

The UWA Institute of Agriculture, The University of Western Australia, Perth, WA, Australia.

Peanut ( L.) is a globally important oil crop, which often experiences poor growth and seedling necrosis under low nocturnal temperatures (LNT). This study assessed the effects of supplementary calcium (Ca) and a calmodulin inhibitor on peanut growth and photosynthetic characteristics of plants exposed to LNT, followed by recovery at a higher temperature. We monitored key growth and photosynthetic parameters in a climate-controlled chamber in pots containing soil. LNT reduced peanut growth and dry matter accumulation, enhanced leaf nonstructural carbohydrates concentrations and non-photochemical quenching, decreased the electron transport rate, increased the transmembrane proton gradient, and decreased gas exchange rates. In peanuts subjected to LNT, foliar application of Ca restored growth, dry matter production and leaf photosynthetic capacity. In particular, the foliar Ca application restored temperature-dependent photosynthesis feedback inhibition due to improved growth/sink demand. Foliar sprays of a calmodulin inhibitor further deteriorated the effects of LNT which validated the protective role of Ca in facilitating LNT tolerance of peanuts.
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http://dx.doi.org/10.3389/fpls.2019.01637DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6985363PMC
January 2020

Cyclic electron flow modulate the linear electron flow and reactive oxygen species in tomato leaves under high temperature.

Plant Sci 2020 Mar 24;292:110387. Epub 2019 Dec 24.

College of Horticulture, Shenyang Agricultural University, Shenyang, 110866, China; Key Laboratory of Protected Horticulture of Education Ministry and Liaoning Province, Shenyang, 110866, China; Collaborative Innovation Center of Protected Vegetable Surrounds Bohai Gulf Region, Shenyang, 110866, China. Electronic address:

The cyclic electron flow (CEF) around photosystem I (PSI) plays a crucial role in photosynthesis and also functions in plant tolerance of abiotic environmental stress. However, the role of PGR5/PGRL1- and NDH-dependent CEF in tomato under hightemperature (HT) is poorly understood. Here, we assessed the photoprotective effect of these pathways in tomato leaves under HT by using antimycin A (AA) and rotenone (R), which are chemical inhibitors of PGR5/PGRL1- and NDH-dependent CEF, respectively. The results showed that AA treatment caused significantly greater inhibition of CEF under HT compared to R treatment. Moreover, AA treatment caused a greater decrease in maximal photochemistry efficiency (F/F) and increased damage to the donor and acceptor side of photosystem II (PSII); however, the limitation of the acceptor side in PSI [Y(NA)] was significantly increased. In addition, thylakoid membrane integrity was compromised and reactive oxygen species, proton gradient (ΔpH), antioxidant enzyme activity, and the expression of photosystem core subunit genes were significantly decreased under AA treatment. These findings indicate that PGR5/PGRL1-dependent CEF protects PSII and PSI from photooxidative damage through the formation of ΔpH while maintaining thylakoid membrane integrity and normal gene expression levels of core photosystem components. This study demonstrates that PGR5/PGRL1-dependent CEF plays a major role in HT response in tomato.
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http://dx.doi.org/10.1016/j.plantsci.2019.110387DOI Listing
March 2020

Effects of Elevated Root-Zone CO on Root Morphology and Nitrogen Metabolism Revealed by Physiological and Transcriptome Analysis in Oriental Melon Seedling Roots.

Int J Mol Sci 2020 Jan 25;21(3). Epub 2020 Jan 25.

College of Horticulture, Shenyang Agricultural University, Shenyang 110866, China.

Rhizosphere CO is vital for crop growth, development, and productivity. However, the mechanisms of plants' responses to root-zone CO are unclear. Oriental melons are sensitive to root-zone gas, often encountering high root-zone CO during cultivation. We investigated root growth and nitrogen metabolism in oriental melons under T1 (0.5%) and T2 (1.0%) root-zone CO concentrations using physiology and comparative transcriptome analysis. T1 and T2 increased root vigor and the nitrogen content in the short term. With increased treatment time and CO concentration, root inhibition increased, characterized by decreased root absorption, incomplete root cell structure, accelerated starch accumulation and hydrolysis, and cell aging. We identified 1280 and 1042 differentially expressed genes from T1 and T2, respectively, compared with 0.037% CO-grown plants. Among them, 683 co-expressed genes are involved in stress resistance and nitrogen metabolism (enhanced phenylpropanoid biosynthesis, hormone signal transduction, glutathione metabolism, and starch and sucrose metabolism). Nitrogen metabolism gene expression, enzyme activity, and nitrogen content analyses showed that short-term elevated root-zone CO mainly regulated plant nitrogen metabolism post-transcriptionally, and directly inhibited it transcriptionally in the long term. These findings provided a basis for further investigation of nitrogen regulation by candidate genes in oriental melons under elevated root-zone CO.
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http://dx.doi.org/10.3390/ijms21030803DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7037942PMC
January 2020

Expression of galactinol synthase from Ammopiptanthus nanus in tomato improves tolerance to cold stress.

J Exp Bot 2020 01;71(1):435-449

Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, PR China.

Soluble carbohydrates not only directly affect plant growth and development but also act as signal molecules in processes that enhance tolerance to cold stress. Raffinose family oligosaccharides (RFOs) are an example and play an important role in abiotic stress tolerance. This study aimed to determine whether galactinol, a key limiting factor in RFO biosynthesis, functions as a signal molecule in triggering cold tolerance. Exposure to low temperatures induces the expression of galactinol synthase (AnGolS1) in Ammopiptanthus nanus, a desert plant that survives temperatures between -30 °C to 47 °C. AnGolS1 has a greater catalytic activity than tomato galactinol synthase (SlGolS2). Moreover, SlGolS2 is expressed only at low levels. Expression of AnGolS1 in tomato enhanced cold tolerance and led to changes in the sugar composition of the seeds and seedlings. AnGolS1 transgenic tomato lines exhibited an enhanced capacity for ethylene (ET) signaling. The application of galactinol abolished the repression of the ET signaling pathway by 1-methylcyclopropene during seed germination. In addition, the expression of ERF transcription factors was increased. Galactinol may therefore act as a signal molecule affecting the ET pathway.
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http://dx.doi.org/10.1093/jxb/erz450DOI Listing
January 2020

Multi-phase level set algorithm based on fully convolutional networks (FCN-MLS) for retinal layer segmentation in SD-OCT images with central serous chorioretinopathy (CSC).

Biomed Opt Express 2019 Aug 15;10(8):3987-4002. Epub 2019 Jul 15.

Shandong Key Laboratory of Medical Physics and Image Processing, Shandong Provincial Engineering and Technical Center of Light Manipulation, School of Physics and Electronics, Shandong Normal University, Jinan, 250014, China.

As a function of the spatial position of the optical coherence tomography (OCT) image, retinal layer thickness is an important diagnostic indicator for many retinal diseases. Reliable segmentation of the retinal layer is necessary for extracting useful clinical information. However, manual segmentation of these layers is time-consuming and prone to bias. Furthermore, due to speckle noise, low image contrast, retinal detachment, and also irregular morphological features make the automatic segmentation task challenging. To alleviate these challenges, in this paper, we propose a new coarse-fine framework combining the full convolutional network (FCN) with a multiphase level set (named FCN-MLS) for automatic segmentation of nine boundaries in retinal spectral OCT images. In the coarse stage, FCN is used to learn the characteristics of specific retinal layer boundaries and achieve classification of four retinal layers. The boundaries are then extracted and the remaining boundaries are initialized based on a priori information about the thickness of the retinal layer. In order to prevent the overlapping of the segmentation interfaces, a regional restriction technique is used in the multi-phase level to evolve the boundaries to achieve fine nine retinal layers segmentation. Experimental results on 1280 B-scans show that the proposed method can segment nine retinal boundaries accurately. Compared with the manual delineation, the overall mean absolute boundary location difference and the overall mean absolute thickness difference were 5.88 ± 2.38μm and 5.81 ± 2.19μm, which showed a good consistency with manual segmentation by the physicians. Our experimental results also outperform state-of-the-art methods.
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http://dx.doi.org/10.1364/BOE.10.003987DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6701532PMC
August 2019

Targeted Control of Chloroplast Quality to Improve Plant Acclimation: From Protein Import to Degradation.

Front Plant Sci 2019 25;10:958. Epub 2019 Jul 25.

Key Laboratory of Protected Horticulture of Ministry of Education, National & Local Joint Engineering Research Center of Northern Horticultural Facilities Design & Application Technology (Liaoning), Horticulture Department, Shenyang Agricultural University, Shenyang, China.

The chloroplast is an important energy-producing organelle acting as an environmental sensor for the plant cell. The normal turnover of the entire damaged chloroplast and its specific components is required for efficient photosynthesis and other metabolic reactions under stress conditions. Nuclear-encoded proteins must be imported into the chloroplast through different membrane transport complexes, and the orderly protein import plays an important role in plant adaptive regulation. Under adverse environmental conditions, the damaged chloroplast or its specific components need to be degraded efficiently to ensure normal cell function. In this review, we discuss the molecular mechanism of protein import and degradation in the chloroplast. Specifically, quality control of chloroplast from protein import to degradation and associated regulatory pathways are discussed to better understand how plants adapt to environmental stress by fine-tuning chloroplast homeostasis, which will benefit breeding approaches to improve crop yield.
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http://dx.doi.org/10.3389/fpls.2019.00958DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6670758PMC
July 2019

Low Overnight Temperature-Induced Gibberellin Accumulation Increases Locule Number in Tomato.

Int J Mol Sci 2019 Jun 21;20(12). Epub 2019 Jun 21.

College of Horticulture, Shenyang Agricultural University, National & Local Joint Engineering Research Center of Northern Horticultural Facilities Design & Application Technology (Liaoning), Key Laboratory of Protected Horticulture of Education Ministry and Liaoning Province, Shenyang 110866, China.

The number of locules in tomato affects fruit size, shape, and the incidence of malformation. Low temperature increases locule number and the incidences of malformation in tomato plants. In this study, three flower bud developmental stages (pre-flower bud differentiation, sepal and petal primordium formation, and carpel primordium formation) under different night temperatures (10, 15, and 20 °C) were used to analyze the reason behind locule number change using an RNA sequencing (RNA-seq) approach, Quantitative real-time PCR (qRT-PCR), and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS). The results showed that the "plant hormone signal transduction", "starch and sucrose metabolism", and "diterpenoid biosynthesis" categories were remarkably activated during flower bud differentiation. Transcripts of gibberellin (GA)-related genes and endogenous levels of GAs were analyzed, and it was discovered that genes were significantly downregulated and bioactive GA and GA accumulated at lower overnight temperature. Exogenous application of bioactive GA, GA, and PAC (paclobutrazol) showed that GA and GA increased the locule number, while PAC decreased the locule number. Taken together, our results suggest that lower overnight temperature reduced the expression of genes, leading to GA and GA accumulation, thereby increasing locule number in tomato.
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http://dx.doi.org/10.3390/ijms20123042DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6627202PMC
June 2019

A family of auxin conjugate hydrolases from Solanum lycopersicum and analysis of their roles in flower pedicel abscission.

BMC Plant Biol 2019 Jun 3;19(1):233. Epub 2019 Jun 3.

Horticulture Department, Shenyang Agricultural University, No. 120 Dongling Road, Shenhe District, Shenyang, 110866, Liaoning, China.

Background: Auxin conjugates are hydrolyzed to release free auxin to ensure defined cellular auxin levels or gradients within tissues for proper development or response to environmental signals. The auxin concentration in the abscission zone (AZ) is thought to play an important role in mediating the abscission lag phase.

Results: In this study, the full cDNA sequences of seven tomato ILR1-like SlILL genes were identified and characterized, All SlILLs were found to have auxin conjugate hydrolysis activity. The effects of different auxin conjugates on abscission identified IAA-Ile as a candidate to determine the auxin conjugate and auxin conjugate hydrolysis functions in abscission. Treatment of pedicel explants with IAA-Ile for different times showed that application before 6 h could effectively delay abscission. IAA-Ile pre-incubation for 2 h was sufficient to inhibit abscission. These results showed that there is not sufficient auxin conjugates in the AZ to inhibit abscission, and the optimal time to inhibit abscission by the application of exogenous auxin conjugates is before 6 h. Treatment with cycloheximide (CHX, a protein biosynthesis inhibitor) indicated that de novo synthesis of auxin conjugate hydrolases is also required to delay abscission. During abscission, SlILL1, 5, and 6 showed abscission-related gene expression patterns, and SlILL1, 3, 5, 6, and 7 showed increasing expression trends, which collectively might contribute to delay abscission. Silencing the expression of SlILL1, 3, 5, 6, and 7 using virus-induced gene silencing showed that SlILL1, 5, and 6 are major mediators of abscission in tomato.

Conclusions: In the process of abscission, auxin inhibition is concentration dependent, and the concentration of auxin in the AZ was regulated by hydrolyzed auxin conjugates. SlILR1, 5, and 6 play a key role in flower pedicel abscission.
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http://dx.doi.org/10.1186/s12870-019-1840-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6547480PMC
June 2019

Rhodamine B in spices determined by a sensitive UPLC-MS/MS method.

Food Addit Contam Part B Surveill 2019 Mar 21;12(1):59-64. Epub 2018 Nov 21.

a Physical and Chemical Laboratory , Center for Disease Control and Prevention of Shaanxi Province , Xi'an , China.

Rhodamine B (RhB) is a banned food additive and has been classified as illegal colourant. Therefore, the risk of RhB contamination should be strictly monitored. In this study, a sensitive UPLC-MS/MS method was applied to monitor RhB in 292 various spices such as chilli, pepper and tomato products. The results showed 22.7% of chilli powder samples, 18.5% of pepper powder samples, 11.1% of chilli oil samples and 9.1% of pepper oil samples were contaminated with RhB. Chilli powder contained RhB up to 44,935 μg/kg with an average of 743 μg/kg, pepper powder up to 65.9 μg/kg with an average of 4.1 μg/kg, chilli oil up to 14.6 μg/kg with an average of 1.0 μg/kg and pepper oil up to 1.1 μg/kg with an average of 0.2 μg/kg, respectively. Considering the common consumption of chilli products and pepper products by so many consumers, RhB exposure is significant and should be decreased.
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http://dx.doi.org/10.1080/19393210.2018.1548504DOI Listing
March 2019

[Assessment on exposure of dietary aluminium among residents in Shaanxi Province during 2013-2015].

Wei Sheng Yan Jiu 2018 Mar;47(2):307-311

Physical and Chemical Laboratory, Shaanxi Center for Disease Control and Prevention, Xi'an 710054, China.

Objective: To investigate the contamination condition of the aluminium in commercial foods in Shaanxi Province, and evaluate aluminium dietary intake level in Shaanxi population and its potential health risks.

Methods: 1331 samples in Shaanxi Province were collected from 2013 to 2015. The aluminium contents were detected by National Food Contamination and Harmful Factors in Risk Monitoring Manual, and data of total diet study in Shaanxi Province in 2007, to calculate dietary intake of aluminium in Shaanxi population.

Results: Average dietary intake of aluminium was 0. 358 mg/( kg·d), accounted for 125% of PTWI. The dietary intake of aluminium in children aged 2 to7 years old was highest, followed children aged 8 to 12 years old and men aged 13 to 19 years old. Grain products and potato products were the main sources of aluminium exposure compared with others.

Conclusion: The dietary intake of aluminium in Shaanxi population is more than the PTWI. The children aged 2 to 7 years old, children aged 8 to12 years old and men aged 13 to 19 years old is high. Grain products and potato products are the main sources of aluminium exposure in Shaanxi population. The effective measureto control the intake of aluminium is to normalize and rationalize the use of additives containing aluminium.
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March 2018

MicroRNA1917 targets CTR4 splice variants to regulate ethylene responses in tomato.

J Exp Bot 2018 02;69(5):1011-1025

College of Horticulture, Shenyang Agricultural University, Shenyang, China.

Ethylene perception is regulated by receptors, and the downstream protein CONSTITUTIVE TRIPLE RESPONSE1 is a key suppressor of ethylene signalling. The non-conserved tomato (Solanum lycopersicum) microRNA1917 (Sly-miR1917) mediates degradation of SlCTR4 splice variants (SlCTR4sv) but the molecular details of this pathway remain unknown. Sly-miR1917 and the targeted SlCTR4sv are ubiquitously expressed in all tomato organs. Overexpression of Sly-miR1917 enhances ethylene responses, including the triple response in etiolated seedlings, in the absence of ethylene, as well as epinastic petiole growth, accelerated pedicel abscission, and fruit ripening. Enhanced ethylene signalling in Sly-miR1917-overexpressing plants (1917-OE) is accompanied by up-regulation of ethylene biosynthesis and signalling genes, and increased ethylene emission. These phenotypes were recovered by repressing the positive ethylene regulator EIN2. Moreover, the Sly-miR1917-targeted SlCTR4 splice variant SlCTR4sv3, expressed specifically in the abscission zone, exhibited the opposite expression pattern to Sly-miR1917. Complementation of the Arabidopsis thaliana ctr-1 mutant and yeast two-hybrid and bimolecular fluorescence complementation assays suggested that SlCTR4sv3 functions in ethylene signalling. Co-expression of Sly-miR1917 and SlCTR4sv3 in Nicotiana benthamiana further suggested that Sly-miR1917 cleaves SlCTR4sv3 in vivo. Database homology searching revealed a Solanum tuberosum CTR-like splice variant containing a Sly-miR1917 binding sequence, and a homologue of mature Sly-miR1917 in potato, indicating a conserved function for miR1917 and the regulatory miRNA-mediated ethylene network in solanaceous species.
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http://dx.doi.org/10.1093/jxb/erx469DOI Listing
February 2018

SlPIN1 regulates auxin efflux to affect flower abscission process.

Sci Rep 2017 11 2;7(1):14919. Epub 2017 Nov 2.

College of Horticulture, Shenyang Agricultural University, Shenyang, 110866, Liaoning, P. R. China.

Solanum lycopersicum PIN-FORMED1 (SlPIN1), a major auxin efflux facilitator, contributes to the establishment of auxin maxima during organ initiation and development in tomato. However, the functions of SlPIN1 during organ abscission remain unclear. In our study, SlPIN1 expression decreased immediately after flower removal and increased following IAA treatment, indicating a high sensitivity to auxin depletion. 1-MCP (an ethylene inhibitor) delayed abscission and down-regulated SlPIN1, indicating that ethylene may positively regulate SlPIN1 and that low expression levels of SlPIN1 may delay abscission. The SlPIN1 protein levels were not consistent with the expression pattern, implying that in addition to transcription, protein degradation also affects SlPIN1 levels during abscission. The phosphorylation of SlPIN1 at Ser418, which significantly declined during abscission, was found to play roles in SlPIN1 localization and auxin transport. We also identified the interaction proteins of SlPIN1, which were involved in phosphorylation and ubiquitylation. Therefore, complex mechanisms mediate SlPIN1 auxin transport capability during abscission. The silencing of SlPIN1 expression accelerated abscission by increasing auxin accumulation in the ovary and decreasing the auxin content in the abscission zone (AZ), indicating that SlPIN1 plays a major role in mediating auxin source-sink transport and the establishment and maintenance of auxin maxima in the AZ.
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http://dx.doi.org/10.1038/s41598-017-15072-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5668252PMC
November 2017

Tomato Transcription Factor Plays an Important Role in Tomato Flower and Locule Development.

Front Plant Sci 2017 31;8:457. Epub 2017 Mar 31.

Key Laboratory of Protected Horticulture, Ministry of Education, Shenyang Agricultural UniversityShenyang, China.

Tomato is a model species for fleshy fruit development. The shapes and sizes of tomato () are mainly controlled by several loci, including locule number (). Two single nucleotide polymorphisms were found downstream of () in a putative tomato CArG -regulatory element. The mutation may affect the binding of (), cause the up-regulation of and result in increased locule numbers. In this study, tissue expression levels showed that is expressed in young floral buds and shoot apexes. Silencing on an mutant genetic background with an RNA interference (RNAi) strategy resulted in smaller flowers and fruit than those of the wild-type plants, with decreased locule number. Further study revealed that the RNAi lines exhibited altered expression levels of the and genes that participate in the regulation of tomato flower and fruit locule development. In conclusion, this study provides the first genetic evidence that may be the candidate gene of the locus and reveals the function of in flower development.
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http://dx.doi.org/10.3389/fpls.2017.00457DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374213PMC
March 2017

Sub-high Temperature and High Light Intensity Induced Irreversible Inhibition on Photosynthesis System of Tomato Plant ( L.).

Front Plant Sci 2017 16;8:365. Epub 2017 Mar 16.

College of Horticulture, Shenyang Agricultural UniversityShenyang, China; Key Laboratory of Protected Horticulture of Education Ministry and Liaoning ProvinceShenyang, China; Collaborative Innovation Center of Protected Vegetable Surrounds Bohai Gulf RegionShenyang, China.

High temperature and high light intensity is a common environment posing a great risk to organisms. This study aimed to elucidate the effects of sub-high temperature and high light intensity stress (HH, 35°C, 1000 μmol⋅m⋅s) and recovery on the photosynthetic mechanism, photoinhibiton of photosystem II (PSII) and photosystem I (PSI), and reactive oxygen (ROS) metabolism of tomato seedlings. The results showed that with prolonged stress time, net photosynthetic rate (Pn), Rubisco activity, maximal photochemistry efficiency (Fv/Fm), efficient quantum yield and electron transport of PSII [Y(II) and ETR(II)] and PSI [Y(I) and ETR(I)] decreased significantly whereas yield of non-regulated and regulated energy dissipation of PSII [Y(NO) and Y(NPQ)] increased sharply. The donor side limitation of PSI [Y(ND)] increased but the acceptor side limitation of PSI [Y(NA)] decreased. Content of malondialdehyde (MDA) and hydrogen peroxide (HO) were increased while activity of superoxide dismutase (SOD) and peroxidase (POD) were significantly inhibited compared with control. HH exposure affected photosynthetic carbon assimilation, multiple sites in PSII and PSI, ROS accumulation and elimination of L.
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http://dx.doi.org/10.3389/fpls.2017.00365DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352666PMC
March 2017
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